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Carbohydrate structural isomers analyzed by sequential mass spectrometry.

David J Ashline1, Anthony J Lapadula, Yan-Hui Liu

  • 1The Glycomics Center, Department of Biochemistry and Molecular Biology, University of New Hampshire, Durham, New Hampshire 03824, USA.

Analytical Chemistry
|April 3, 2007
PubMed
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This study introduces advanced mass spectrometry (MSn) methods for characterizing carbohydrate structural isomers, crucial for understanding complex biological structures like those in ovalbumin and IgG.

Area of Science:

  • Glycomics
  • Analytical Chemistry
  • Structural Biology

Background:

  • Comprehensive carbohydrate sequencing is essential for understanding biological functions.
  • Characterizing structural isomers of carbohydrates presents significant analytical challenges.
  • Previous methods lacked the resolution to fully elucidate complex glycan structures.

Purpose of the Study:

  • To develop and validate advanced mass spectrometry (MSn) protocols for the characterization of carbohydrate structural isomers.
  • To apply these methods to identify and analyze previously unreported glycans.
  • To establish a robust strategy for carbohydrate sequencing and isomer resolution.

Main Methods:

  • Utilized ion trap instrumentation for sequential mass spectrometry (MSn) with automation and computational tools.

Related Experiment Videos

  • Developed protocols based on the principle of reproducible fragmentation spectra for structural elucidation.
  • Employed gas-phase isolation and fragmentation of anomalous ions to resolve specific isomers.
  • Incorporated derivatization techniques to aid in substructure positioning within glycan arrays.

    • Main Results:

    • Successfully characterized structural isomers of carbohydrates, including those in ovalbumin and IgG.
    • Identified several previously unreported glycans in IgG samples.
    • Demonstrated that multiple stages of mass spectrometry (MSn>2) are sometimes required for isomer characterization.
    • Bioinformatics tools were effective in assimilating structural data from MSn datasets.

    Conclusions:

    • Advanced MSn techniques provide a powerful strategy for resolving and characterizing carbohydrate structural isomers.
    • The developed methods enhance the comprehensive sequencing of complex carbohydrates.
    • This approach significantly advances the field of glycomics by enabling the discovery of novel glycan structures.